骨形态发生蛋白-6对转化生长因子-β2所致视网膜色素上皮细胞迁移的影响  

Effects of bone morphogenetic protein-6 on the migration of retinal pigment epithelial cells induced by transforming growth factor-β2

作　　者：吉梦 刘明[3] 马波[1] 刘轩[1] 裴澄[1] 陈丽[1] JI Meng;LIU Ming;MA Bo;LIU Xuan;PEI Cheng;CHEN Li(Department of Ophthalmology,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China;Department of Ophthalmology,Ankang Hospital of Traditional Chinese Medicine,Ankang Eye Hospital,Ankang 725000,Shaanxi Province,China;Department of Ophthalmology,Xi’an No.1 Hospital,Shaanxi Institute of Ophthalmology,Xi’an 710001,Shaanxi Province,China)

机构地区：[1]西安交通大学医学院第一附属医院眼科,陕西省西安市710061 [2]安康市中医医院眼科,安康市眼科医院,陕西省安康市725000 [3]西安市第一医院眼科,陕西省眼科研究所,陕西省西安市710001

出　　处：《眼科新进展》2021年第8期717-721,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助项目(编号81800812);陕西省自然科学基金资助项目(编号2020SF268)。

摘　　要：目的探讨骨形态发生蛋白-6(BMP-6)对转化生长因子-β2(TGF-β2)所致视网膜色素上皮(RPE)细胞迁移的影响。方法RPE细胞常规培养后,用5μg·L-1 TGF-β2处理48 h,Western blot检测细胞中BMP-6蛋白水平的变化,Transwell法检测细胞迁移能力变化。随后,筛选BMP-6 siRNA抑制率最高的片段用于后续实验,构建BMP-6过表达质粒并进行鉴定后用于后续实验,RPE细胞随机分组,即(1)对照组加入正常培养基培养48 h;(2)TGF-β2组培养基中加入5μg·L-1 TGF-β2培养48 h;(3)BMP-6 siRNA组转染BMP-6-homo-1087后培养48 h;(4)BMP-6过表达组转染BMP-6过表达质粒后培养48 h;(5)TGF-β2+BMP-6过表达组转染BMP-6过表达质粒24 h后,加入5μg·L-1TGF-β2继续培养48 h。Transwell法检测细胞迁移数的变化,倒置显微镜观察细胞形态的变化。结果TGF-β2组迁移细胞数为(122.00±5.57)个,与对照组(63.67±4.04)个相比差异有统计学意义(P=0.000)。TGF-β2组BMP-6蛋白表达水平为0.41±0.13,与对照组(0.70±0.08)相比差异具有统计学意义(P=0.031)。BMP-6 siRNA组迁移细胞数为(115.67±1.53)个,与对照组(57.00±7.00)个相比差异具有统计学意义(P=0.000)。BMP-6过表达组迁移细胞数为(44.33±5.51)个,与对照组(60.00±6.25)个相比差异无统计学意义(P=0.076)。TGF-β2+BMP-6过表达组迁移细胞数为(90.33±3.51)个,与TGF-β2组(112.00±9.64)个相比差异具有统计学意义(P=0.016)。结论BMP-6可以阻止TGF-β2所致的RPE细胞的迁移,为增生性玻璃体视网膜病变的防治提供新的视角。Objective To investigate the effect of bone morphogenetic protein-6(BMP-6)on the migration of retinal pigment epithelial(RPE)cells induced by the transforming growth factor-β2(TGF-β2).Methods After being routinely cultured,RPE cells were treated with 5μg·L-1 TGF-β2 for 48 h.Their BMP-6 protein levels were detected by the Western blot,and cell migration was detected by the Transwell chamber.Subsequently,the fragment with the highest inhibition rate of BMP-6 siRNA was selected for subsequent experiments,and BMP-6 overexpression plasmids were constructed and identified for use in subsequent experiments.RPE cells were randomly divided into(1)control group cultured with the normal medium for 48 h;(2)TGF-β2 group cultured with 5μg·L-1 TGF-β2 for 48 h;(3)BMP-6 siRNA group transfected with the BMP6-homo-1087 and cultured for 48 h;(4)BMP-6 overexpression group transfected with the BMP-6 overexpression plasmid and cultured for 48 h;(5)TGF-β2+BMP-6 overexpression group after transfection with the BMP-6 overexpression plasmid for 24 h,cultured for 48 h with additional 5μg·L-1 TGF-β2.The Transwell chamber was used to detect the change in the number of migrating cells,and the inverted microscope to observe the change in the cell morphology.Results The number of migrating cells in the TGF-β2 group was 122.00±5.57,which was significantly higher than that in the control group(63.67±4.04)(P=0.000).The expression level of BMP-6 protein in the TGF-β2 group was 0.41±0.13,which was significantly lower than that in the control group(0.70±0.08)(P=0.031).The number of migrating cells in the BMP-siRNA group was 115.67±1.53,which was significantly higher than that in the control group(57.00±7.00)(P=0.000).The number of migrating cells in the BMP-6 overexpression group was 44.33±5.51,which had significant no difference when compared with the control group(60.00±6.25)(P=0.076).The number of migrating cells in the TGF-β2+BMP-6 overexpression group was 90.33±3.51,which was significantly lower than that in the TGF-β2 g

关 键 词：骨形态发生蛋白-6 转化生长因子-Β2 视网膜色素上皮细胞 增生性玻璃体视网膜病变 细胞迁移 

分 类 号：R774[医药卫生—眼科]